Lambda Antibody Light Chain Expression via Engineered Signal Peptides
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Solution Overview
Problem
Current monoclonal antibody production methods face challenges in achieving efficient secretion and cleavage heterogeneity due to non-specific cleavage of signal peptides, leading to variations in antibody yields and potential truncation of N-termini, which can affect the affinity and stability of the final product.
Innovation Solution
The use of specific signal peptides, such as the native and non-native signal peptides for λ light chain variable (VL) domains, to control the cleavage site and N-terminal sequence of the VL domain, ensuring consistent expression and restoring functional activity by engineering the N-terminal sequence to match that of the native leader.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If non-specific signal peptides are used for antibody expression, then antibody yields may increase, but cleavage heterogeneity occurs leading to N-terminal truncation and reduced product quality
Solution Approach 1:
The patent applies parameter changes by modifying the signal peptide sequence to create an engineered signal peptide with optimized cleavage properties. Specifically, the signal peptide is engineered to contain a preferred cleavage site motif (such as Ala-Arg-Cys) that ensures consistent cleavage by signal peptidase, thereby resolving the contradiction between achieving high antibody yields and maintaining cleavage site consistency.
Solution Approach 2:
The engineered signal peptide acts as an intermediary element that mediates between the antibody polypeptide and the cellular secretion machinery. By designing the signal peptide with specific structural features and cleavage motifs, it ensures proper translocation into the endoplasmic reticulum followed by precise cleavage, thus maintaining manufacturing precision while enabling efficient secretion and high productivity.
2Productivity
If signal peptides are used to direct translocation into the endoplasmic reticulum, then antibody secretion efficiency improves, but cleavage heterogeneity leads to elongation or truncation of N-termini
Solution Approach 1:
The patent modifies the signal peptide parameters by engineering specific amino acid sequences that create a preferred cleavage site. The engineered signal peptide contains motifs such as Ala-Arg-Cys at its C-terminus, which are recognized by signal peptidase for precise cleavage. This parameter change ensures that while the signal peptide efficiently directs translocation into the endoplasmic reticulum (improving secretion efficiency), it also ensures consistent cleavage at a specific site (maintaining N-terminal sequence consistency and reliability).
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach results in consistent and efficient expression of λ antibodies with desired properties, improving antibody yields and stability by controlling the N-terminal sequence, thereby enhancing the functional activity and reducing heterogeneity.
Implementation Method 1
translocation of the antibody polypeptides into the lumen of the endoplasmic reticulum (ER), via signal peptides
Implementation Method 2
The signal peptide is a short (15-30 amino acid) sequence at the N terminus of the antibody heavy and light chains, which directs its translocation and is cleaved during the translocation process
Data Source
AI summary
Expression of lambda antibody light chain with non-native leader sequence, wherein light chain is sequence-engineered to restore native N terminus. Immunoglobulin lambda variable domain sequence comprising N terminal deletion for expression with non-native N terminal signal peptide.